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| P1 of strawberry vein banding virus, a multilocalized protein, functions as a movement protein and interacts with the coat protein |
| RUI Peng-huan1*, WANG Zhan-qi2*, SHAN Wen-shu1*, XIA Wei-wei1, ZHOU Xiu-hong3, YANG Lian-lian2, JIANG Lei1, 4, 5, JIANG Tong1, 4, 5 |
1 School of Plant Protection, Anhui Agricultural University, Hefei 230036, P.R.China
2 Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Province, College of Life Sciences, Huzhou University, Huzhou 313000, P.R.China
3 Biotechnology Center, Anhui Agricultural University, Hefei 230036, P.R.China
4 Anhui Province Key Laboratory of Integrated Pest Management on Crops, Hefei 230036, P.R.China
5 Key Laboratory of Biology and Sustainable Management of Plant Diseases and Pests of Anhui Higher Education Institutes, Anhui Agricultural University, Hefei 230036, P.R.China |
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摘要
尽管草莓镶脉病毒（SVBV）的基因组全序列已经确定，生物信息学分析显示SVBV基因组可编码7个不同的蛋白，但每个蛋白的确切功能并不是非常清楚。在本研究中，我们提供了如下能够验证SVBV编码的P1蛋白（SVBV-P1）所具有的几个特点的证据。利用蛋白定位预测工具分析SVBV-P1蛋白亚细胞定位，共聚焦观察SVBV-P1浸润的本氏烟叶片，结果显示SVBV-P1蛋白定位于本氏烟表皮细胞的细胞质、细胞壁和胞间连丝，还能够在细胞质中形成与微管和内质网相关的包涵体。稀释的SVBV-P1浸润本氏烟叶片，使SVBV-P1蛋白在单细胞表达，结果显示一段时间之后SVBV-P1蛋白能从本氏烟叶片上农杆菌浸润的侵染点细胞转移到邻近的细胞，表明SVBV-P1蛋白具有胞间移动的能力。进一步将稀释的SVBV-P1与马铃薯X病毒（PVX）运动缺陷型突变体共浸润本氏烟叶片，结果显示PVX突变体能从单细胞移动至周围的细胞，表明SVBV-P1蛋白能促进失去运动功能的PVX突变体在本氏烟叶片的细胞间运动。最后，酵母双杂交实验与双分子荧光互补分析显示，SVBV-P1与SVBV-CP共转化的酵母菌能正常生长，SVBV-P1与SVBV-CP共浸润的本氏烟叶片出现荧光，表明SVBV-P1蛋白能与SVBV编码的外壳蛋白互作，而外壳蛋白是花椰菜花叶病毒属病毒粒子的主要成分。电泳迁移率试验测定结果显示，与SVBV-P1蛋白孵育的DNA在电泳过程中并未出现阻滞，说明SVBV-P1蛋白缺乏结合DNA的能力。综上所述，我们的研究结果表明SVBV-P1蛋白很可能是SVBV的运动蛋白，并且为进一步研究花椰菜花叶病毒属病毒的运动蛋白功能提供了新的思路。
Abstract Although the complete nucleotide sequence of strawberry vein banding virus (SVBV) has been determined and bioinformatic analysis has revealed that the SVBV genome could encode seven proteins, the precise function of each protein is unclear. This study provided evidence that the P1 protein of SVBV (SVBV-P1) possesses the following features. Bioinformatic and subcellular localization analyses showed that SVBV-P1 is localized in the cytoplasm and cell walls of epidermal cells in Nicotiana benthamiana, and it forms inclusion bodies associated with microtubules and the endoplasmic reticulum. Dilution experiments demonstrated that SVBV-P1 could move from the original agro-infiltrated cells to adjacent cells in N. benthamiana leaves. Further trans-complementation experiments demonstrated that SVBV-P1 could facilitate the intercellular movement of a movement-deficient potato virus X mutant in N. benthamiana leaves. Finally, yeast two-hybrid and bimolecular fluorescence complementation assays revealed that SVBV-P1 could interact with the SVBV coat protein, which is a major component of Caulimovirus virions. Results of the electrophoretic mobility shift assay indicated that SVBV-P1 lacks DNA-binding capability. In summary, the results suggest that SVBV-P1 is probably a movement protein of SVBV, providing new insights into the function of movement proteins of the Caulimovirus genus.
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Received: 06 December 2020
Accepted: 16 April 2021
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| Fund: This work was supported by the grants from the National Natural Science Foundation of China (32072386 and 31801700), the Key Research and Development Project of Anhui Province, China (202004a06020013) and the Postdoctoral Science Fund of Anhui Province, China (2019B360). |
| About author: RUI Peng-huan, E-mail: 15755308130@163.com; WANG Zhan-qi, E-mail: zhqwang@zju.edu.cn; SHAN Wen-shu, E-mail: shanwenshu_ah@163.com; Correspondence JIANG Lei, Tel: +86-551-65786312, E-mail: jianglei062x@ahau.edu.cn; JIANG Tong, Tel: +86-551-65786312, E-mail: jiangtong4650@sina.com
* These authors contributed equally to this study. |
Cite this article:
RUI Peng-huan, WANG Zhan-qi, SHAN Wen-shu, XIA Wei-wei, ZHOU Xiu-hong, YANG Lian-lian, JIANG Lei, JIANG Tong.
2022.
P1 of strawberry vein banding virus, a multilocalized protein, functions as a movement protein and interacts with the coat protein. Journal of Integrative Agriculture, 21(4): 1071-1083.
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